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. Author manuscript; available in PMC: 2019 Dec 15.
Published in final edited form as: J Immunol. 2018 Nov 5;201(12):3731–3740. doi: 10.4049/jimmunol.1800333

Figure 2. In vitro macrophage-mediated hepatocytotoxicity is FcγRI- and FcγRIII-dependent.

Figure 2.

Macrophage:hepatocyte co-cultures consisted of FVB/N hepatocytes and RAW 264.7 macrophages activated by pretreatment with IFN-γ (2.5 ng/mL; 18 hours). Prior to co-culture RAW macrophages were incubated with FcγRI and/or FcγRIII blocking antibody (2 μg per million macrophages) or control IgG. The macrophages were washed with PBS and then added to the hepatocyte co-culture for an 8 hour incubation (1.5×106 cells macrophages and 1.5×105 hepatocytes). Control IgG-treated macrophages mediated hepatocytotoxicity against alloantibody incubated target hepatocytes (49.8±2.7%; n=24). Anti-FcγRI mAb (24.3±4.3%; n=11) and anti-FcγRIII mAb (24.7±4.0%; n=9) treatment significantly blocked macrophage-mediated hepatocytotoxicity (p<0.0001 for both, as denoted by “*”). Combination treatment with both anti-FcγRI and anti-FcγRIII mAbs completely inhibited macrophage-mediated hepatocytotoxicity (3.2±1.0%; n=12, p<0.0001, as denoted by “**”). Data is combined from duplicate experiments.